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Dissolution Concepts and Applications.
Gregory P. Martin and Vivian A. Gray
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ABOUT THE AUTHORS
Gregory P. Martin is president of Complectors Consulting (www.complectors.com), which provides consulting and
training in the area of pharmaceutical analytical chemistry. He may be contacted at greg.martin@complectors.com. Viv-
ian A. Gray has 35 years experience in all aspects of dissolution testing and began a consulting business in 2002, V.A.
Consulting, Inc., in dissolution testing and related areas. She may be contacted at vagray@rcn.com.
Validation of
Dissolution Methods
Dissolution Concepts and Applications provides a
forum for sharing information about topics associ-
ated with in vitro dissolution testing. Our objective
for this feature: useful and practical information
applicable to daily work situations.
Reader comments, questions, and suggestions are
needed to help us fulfill the column objective. Please
send your comments and suggestions to column
coordinators Vivian Gray at vagray@rcn.com or Greg
Martin at greg.martin@complectors.com, or to jour-
nal managing editor Susan Haigney at shaigney@
advanstar.com.
KEY POINTS
The following key points are discussed:
Validation of analytical methods in the pharma-
ceutical industry is a well-known requirement
of current good manufacturing practices, and
dissolution methods fall under this requirement.
The validation exercise for dissolution methods
addresses key areas including product perfor-
mance reproducibility, solutions stability, and
detection of changes in formulation, process, and
product performance on stability.
The approach to method validation must be
based on good science and a defined strategy.
This includes identification of the analytical target
profile and associated modern expectations for
dissolution testing methods.
Traditional analytical performance characteristics
including specificity, accuracy, precision, linearity,
and range are discussed.
Other considerations based on risk evaluation,
including deaeration, volume, use of sinkers, fil-
tration, solution stability, carryover from sam-
pling devices, and automation are also discussed.
Validation of a dissolution method provides docu-
mented evidence that the method will be suitable
for its intended use and serve as needed during
the product lifecycle.
INTRODUCTION
The validation of analytical methods in the phar-
maceutical industry is a well-known requirement of
the current good manufacturing practice guidelines
(CGMPs), and dissolution methods fall under this
requirement. While the current regulatory guidances
on method validation such as United States Pharma-
copeia (USP) general chapters <1225> and <1092>,
and International Conference on Harmonisation
(ICH) guidance Q2 (R1) (1-3) provide valuable infor-
mation, the validation of dissolution methods may
require investigation into analytical performance
characteristics that goes beyond what is included
in the guidance documents. This article discusses
some practical approaches for addressing both the
traditional analytical performance characteristics
and some of those that may be unique to dissolu-
tion methods.
The validation exercise for dissolution methods
actually aims to answer the following questions:
Is the variability of the method satisfactory and
will replicate tests of the same sample result in
profiles that are similar to one another?
Are there any significant analytical solutions sta-
bility issues?
Can the method detect changes in formulation
or the manufacturing process?
Does the method have the capability to detect
changes on stability?
Gregory P. Martin and Vivian A. Gray
gxpandj v t . com JOURNAL OF VALIDATION TECHNOLOGY [AUTUMN 2011] 29
Gregory P. Martin and Vivian A. Gray.
STRATEGY AND APPROACH
As always, the approach to method validation starts
with applying good science. Validation should be an
exercise that builds on a firm foundation. Considering
the lifecycle of a dissolution method, there may be
multiple validation exercises. There is an expectation
that even before validation begins, a well-defined
analytical target profile (4) has been documented
and the method has been developed using sound
practices based on an understanding of technology
and expectations with regard to dissolution (5, 6).
These are useful in establishing the acceptance criteria
that will be used to evaluate the validation exercises.
Additionally, it is useful to carry out experiments to
enhance the understanding of the capabilities of the
dissolution method. These include investigating the
impact of various parameters on dissolution results
such as lot-to-lot variability, the influence of different
analysts or instruments, or the effect of stability. It is
also valuable to have experience with genuine samples
that are characteristic of those that will be tested by
the method and not just lab samples.
ANALYTICAL PERFORMANCE
Traditional analytical performance characteristics
are described in USP general chapter <1225> and in
ICH guidance document Q2. These include specificity,
accuracy, precision, linearity, and range. It is gener-
ally appropriate to consider all of these for dissolu-
tion methods, although the extent to which they are
evaluated may depend on the phase of the project (7).
Specificity
Specificity is an important characteristic that should
be considered early during the method development
process (8). This is because specificity requirements
often drive the runtime for a chromatographic analysis
or the selection of wavelength for a spectrophotomet-
ric analysis. There has been a lot of discussion within
the pharmaceutical community about the specific-
ity requirements for a dissolution method. Keep in
mind that dissolution results are typically reported
as integer values. For this reason, interference from
small peaks, such as impurities or degradations that
are controlled to levels well below 1%, may have no
impact on reported dissolution values. However, it is
important to demonstrate that there is no significant
interference (e.g., >2%) from the dissolution medium,
the placebo, or sample manipulation, such as the
effect of filters.
Accuracy
When assessing accuracy, the goal is to evaluate the
recovery of the active ingredient from the dosage form
in the dissolution medium. These experiments gener-
ally include multiple replicates that may be carried out
at the nominal concentration of 100% dissolved or at
several different concentrations covering the range of
interest. Some chemists perform these experiments in
volumetric flasks while others prefer to perform the
experiment in the vessel, using the actual dissolution
apparatus. Typical acceptance criteria for recovery in
dissolution methods are 97-103%.
Precision
Precision is probably the analytical performance char-
acteristic that generates the most interest and complex-
ity for dissolution methods. Recognize that this actually
incorporates a number of different parameters, which
we will address in increasing levels of magnitude. For
instance, it incorporates the variability from the actual
determination, whether we are talking about injection
precision for high-performance liquid chromatography
(HPLC) or replicate measurements on a UV spectropho-
tometer. Next is the variability associated with testing
multiple replicates of the sample. Usually dissolution
testing is performed on multiples of six dosage forms to
be consistent with the USP staged acceptance criteria.
If the variability associated with these replicates is too
high, it could result in the need to perform additional
testing. Another aspect that can contribute to the vari-
ability is called intermediate precision or ruggedness,
and refers to the effects of testing on different days, by
different analysts, or on different instruments. Some-
times intermediate precision is simulated by perform-
ing robustness testing, in which some of the parameters
that may be anticipated to change are intentionally
varied. Finally, in some cases, it is necessary to evalu-
ate inter-laboratory variability. It is not uncommon,
in the lifecycle of an analytical method, to anticipate
that the testing will be performed by more than one
laboratory. In these cases, it is important to evaluate the
effects of transferring a method from one laboratory to
another. The general expectation is that variability at
all but the early time points is relative standard devia-
tion (RSD) less than 10% (9). Often the variability is
much less than this.
Range and Linearity
The analytical range for the method should include
the entire range of anticipated results from the testing.
30 JOURNAL OF VALIDATION TECHNOLOGY [AUTUMN 2011] i v t home. com
Dissolution Concepts and Applications.
For some methods this may include values as low as
10 or 20% of the concentration of a fully dissolved
product, especially where profiles are being collected
or when the product is an extended release dosage
form. The upper end of the range should include val-
ues that are at least 10 to 20% above the concentra-
tion of a fully dissolved product, and may need to go
even higher in situations in which a large number of
samples are being pulled without medium replace-
ment. Linearity should be demonstrated over the full
range of expected values.
Robustness
With regard to robustness, in addition to the usual
parameters that are normally investigated associated
with the determinative step, several parameters may
be of interest for dissolution methods. These include
the buffer concentration, the pH of the medium, the
temperature of the medium in the vessels, the rota-
tion speed of the apparatus, and the timing of the
samples (10).
Other ConsiderationsRisk Analysis
There are several other parameters that should be
investigated when validating a dissolution method.
A rigorous risk assessment is helpful in identifying
parameters that may be important. Some of these
include deaeration, medium volume, effect of sink-
ers, filter qualification, solutions stability, carryover
during sampling, and impact of automation.
Deaeration. The effect of deaeration is fairly unique
to dissolution testing and the magnitude of the impact
may vary significantly from one dissolution method to
another. Deaeration refers to the process of removing
dissolved gases, primarily air, from the dissolution
medium. This may be accomplished in a variety of
ways, including vacuum filtration, sparging with a less
soluble gas such as helium or passing the medium by
a gas permeable membrane (11). Some products seem
to be unaffected by the presence of gases in the disso-
lution medium while there may be significant affects
with other products. The presence of air bubbles has
been observed on the surface of solitude since forms,
which might slow down the rate of dissolution, and
have also been observed attached to granules of dis-
integrated tablets, causing them to rise to the surface
and potentially increase the dissolution rate. When
deaeration has a significant effect on dissolution results,
it must be carefully controlled. A device for measuring
dissolved oxygen or dissolved gases may be useful in
helping to control deaeration.
Volume. It is important to control the volume
of the dissolution medium because any changes in
volume will affect the concentration of the samples
removed. Many laboratories use a graduated cylinder
to measure the dissolution medium. There is vari-
ability associated both with the measurement in the
graduated cylinder and with the transfer process,
because there may be some liquid that is not trans-
ferred from the graduated cylinder.
Sinkers. Sinkers are sometimes used in dissolution
methods, most often to maintain the dosage form at
the bottom of the vessel but also to prevent dosage
forms from sticking to the vessel. When sinkers are
used, it is important to assure consistency from run
to run (12). This requires that the sinkers be carefully
described in the analytical procedure. When using
commercial sinkers, this can be accomplished by
specifying the manufacture and part number. When
sinkers are fabricated in the laboratory, as is the case
when a simple wire coil is used, including adequate
descriptions will help to minimize run-to-run vari-
ability. Recognize that when a method is being used
for multiple potencies, the same sinker may not be
appropriate for all of the potencies. For instance, larger
capsules may not fit well into a sinker that is opti-
mized for smaller capsules.
Filtration. During validation, it is important
to evaluate the impact of the filters used to clarify
samples. First, it should be demonstrated that the
use of the filter does not cause any interference in the
determinative step. Second, it should also be demon-
strated that there is no loss of active drug during the
filtration process.
Stability. Solution stability should be evaluated
for all analytical methods, but is even more important
for dissolution methods. Because the dissolution test
is normally performed at 37C, the elevated tempera-
ture has the potential to decrease solution stability.
Furthermore, because dissolution testing is normally
carried out in aqueous media, compounds that are
poorly water-soluble may have decreased solution sta-
bility, and the solution stability may be affected by the
pH of the medium. Acceptance criteria for solution
stability vary from lab to lab, and may depend on a
number of factors such as the likelihood of needing
additional testing, but are generally in the range of
1 to 3% loss.
Sampling device carryover. Carryover effects
have been observed when using the same sampling
device for multiple vessels or for multiple time points.
This practice has been used in many laboratories,
gxpandj v t . com JOURNAL OF VALIDATION TECHNOLOGY [AUTUMN 2011] 31
Gregory P. Martin and Vivian A. Gray.
because it is sometimes impractical to use a fresh
sampling device for every vessel and for every time
point. In these situations, it is important to evaluate
any potential carryover effects. Carryover is usually
controlled at less than 2%.
Automation. Automation has been used in a vari-
ety of ways to assist with dissolution testing. This
makes sense because dissolution testing is a time-
consuming test. However, it is important to be aware
that the introduction of automation has the potential
to significantly affect dissolution results (13). It is
beyond the scope of this article to address the subject
comprehensively. It should be sufficient to point out
that dissolution testing is unique in that the concen-
tration of active ingredient in the dissolution vessel
may be changing rapidly during the execution of the
test. Therefore, introduction of automation has sig-
nificant potential to affect the results. It is important
to remember that regulatory agencies, when testing a
commercial product, are likely to use manual proce-
dures, and therefore, a correlation between manual
and automated results should be well characterized.
CONCLUSIONS
The goal of validation of a dissolution method is to
provide documented evidence that the method will
be suitable for its intended use. Validation builds on a
foundation of identified requirements, sound develop-
ment practices, and good science. Risk analysis aids in
the identification of additional parameters that may
need to be evaluated, and the analytical target pro-
file can inform the acceptance criteria for evaluating
whether or not the method meets the goals.
REFERENCES
1. USP, Validation of Compendial Procedures, United
States Pharmacopeia 34 National Formulary 29, General
Chapter 1225, 778-792.
2. USP, The Dissolution Procedure: Development and
Validation, United States Pharmacopeia 34 National
Formulary 29, General Chapter 1092, 624-630.
3. ICH, Validation of Analytical Procedures: Text and
Methodology, Q2 (R1), ICH Harmonised Tripartite
Guideline, Nov. 2005.
4. Nethercote, P., Borman, P., Bennett, T., Martin, G., and
McGregor, P. QbD for Better Method Validation and
Transfer, Pharmaceutical Manufacturing, 9(4), 2010
5. FDA, Guidance for Industry: Dissolution Testing of Immedi-
ate Release Solid Oral Dosage Forms, Food and Drug Ad-
ministration, Center for Drug Evaluation and Research
(CDER), August 1997.
6. Gray, V.A., Zheng, J.Y. and Sesi, N. Chapter 11: In Vitro
Dissolution Testing and Method Development, Formula-
tion and Analytical Development for Low-Dose Oral Drug
Products, Wiley, 2009, 265-280.
7. Boudreau, S.P., Martin, L., McElvain, J, Dowling, T. and
Fields, S.M., Method Validation by Phase of Develop-
ment, An Acceptable Analytical Practice, Pharmaceuti-
cal Technology, Nov. 2004, 54-66.
8. Wang, Q.A., and Gray, V.A., Chapter 15 HPLC in Dis-
solution Testing, Handbook of Pharmaceutical Analysis by
HPLC, Elsevier, 2005, 379-399.
9. FDA, Guidance for Industry Waiver of In Vivo Bioavailability
and Bioequivalence Studies for Immediate-Release Solid Oral
Dosage Forms Based on a Biopharmaceutics Classification
System, Food and Drug Administration Center for Drug
Evaluation and Research (CDER) August 2000.
10. Gray, V.A., Identifying Sources of Error and Variability
in Dissolution Calibration and Sample Testing, Ameri-
can Pharmaceutical Review, 5(2)2002, 8-13.
11. Degenhardt, O.S., Waters, B., Rebelo-Cameirao, A.,
Meyers, A., Bruner, H. and Toltl, N. Comparison of
Effectiveness of Various Deaeration Techniques, Dis-
solution Technologies, Feb 2004, 6-11.
12. Brown, W and Marques, M., Question and Answer Sec-
tion, Dissolution Technologies, May 2007, 33.
13. Fortunato, D., A Compliance Perspective on Dissolu-
tion Method Validation for Immediate Release Solid
Oral Dosage Forms on Automated Instruments, Phar-
maceutical Technology, Sept. 2006. JVT